AMYLOD BETA-PEPTIDE SPECIFIC scFv ANTIBODIES

淀粉样 β 肽特异性 SCFV 抗体

• 批准号: 6937814
• 负责人: GOAR GEVORKIAN
• 金额: $ 9.72万
• 依托单位: UNIVERSITY OF MEXICO, UNAM
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6937814
• 关键词: Alzheimer' s disease; amyloid proteins; antibody; bacterial virus; chemical aggregate; combinatorial chemistry; immunologic substance development /preparation; immunotherapy; molecular cloning; neurotoxicology; passive immunization; peptide library; phage display; protein engineering; protein sequence; protein structure

DESCRIPTION (provided by applicant): Amyloid b-peptide (Ab), the major molecular component of the cerebral amyloid plaques, appears to play a central role in the neuropathology of Alzheimer's disease (AD). Compounds that prevent the formation of Ab aggregates or that selectively destroy these aggregates are attractive candidates for the development of therapeutic reagents for prevention and treatment of Alzheimer's disease. Also, compounds that interfere with the interactions of amyloid precursor protein (APP) with other factors that are involved in directing it into pathological pathway as well as those that are capable to prevent or destroy intraneuronal accumulations of Ab, are of great interest for developing therapeutic molecules for Alzheimer's disease. In this project we are proposing the possible immunological intervention for prevention and treatment of AD applying phage display technology for the construction of the first anti-Ab single chain fragment variable (scFv) antibody library and the selection of individual phage clones expressing Ab-specific scFv antibody, capable of preventing the aggregation of Ab or dissolving the preexisting aggregates, scFv phage display library enriched in anti-Ab antibodies will be constructed using the first strand cDNA synthesized from mRNA of spleen and lymph node cells isolated from mice immunized with Ab. The constructed library as well as a human scFv library will be used in bioselection procedure to isolate Ab-specific scFvs expressed on phage. The amino acid sequences of antibody complementarity-determinig regions (CDR) will be prepared and used as "mini-antibodies". Evaluation of in vitro Ab aggregation and neurotoxicity in the presence of the selected compounds: scFv antibodies expressed on phage or in soluble form and "mini-antibodies" will be performed. The most promising molecules selected in this project will be proposed for further evaluation in animal models to all researchers interested in these studies. If successful, these molecules could be of interest for passive immunization in humans, may be after modification with substances that would increase their blood-brain barrier permeability.

描述（由申请人提供）：淀粉样蛋白b-肽（Ab）是大脑淀粉样斑块的主要分子成分，似乎在阿尔茨海默病（AD）的神经病理学中发挥核心作用。防止抗体聚集体形成或选择性破坏这些聚集体的化合物是开发用于预防和治疗阿尔茨海默病的治疗试剂的有吸引力的候选者。此外，干扰淀粉样前体蛋白 (APP) 与其他参与引导其进入病理途径的因子相互作用的化合物以及能够防止或破坏抗体在神经元内积聚的化合物，对于开发阿尔茨海默氏病的治疗分子非常有意义。在这个项目中，我们提出了预防和治疗AD的可能的免疫干预方法，应用噬菌体展示技术构建第一个抗Ab单链片段可变（scFv）抗体库，并选择表达Ab特异性scFv抗体的单个噬菌体克隆，能够防止Ab聚集或溶解预先存在的聚集体，富含抗Ab抗体的scFv噬菌体展示库将被开发出来。 使用从用抗体免疫的小鼠分离的脾和淋巴结细胞的mRNA合成的第一链cDNA构建。构建的文库以及人 scFv 文库将用于生物选择程序，以分离噬菌体上表达的 Ab 特异性 scFv。抗体互补决定区(CDR)的氨基酸序列将被制备并用作“微型抗体”。将在所选化合物存在下评估体外抗体聚集和神经毒性：在噬菌体上或以可溶形式表达的 scFv 抗体和“微型抗体”。该项目中选择的最有前途的分子将被建议在动物模型中向所有对这些研究感兴趣的研究人员进行进一步评估。如果成功，这些分子可能对人类被动免疫感兴趣，可能会用增加血脑屏障渗透性的物质进行修饰。

项目成果

Identification of amyloid-beta 1-42 binding protein fragments by screening of a human brain cDNA library.

通过筛选人脑 cDNA 文库鉴定淀粉样蛋白 β 1-42 结合蛋白片段。

• DOI: 10.1016/j.neulet.2005.11.061
• 发表时间: 2006
• 期刊: Neuroscience letters
• 影响因子: 2.5
• 作者: Munguia,MariaElena;Govezensky,Tzipe;Martinez,Rodrigo;Manoutcharian,Karen;Gevorkian,Goar
• 通讯作者: Gevorkian,Goar